Project description:Background The number of managed honey bee colonies has considerably decreased in many developed countries in recent years and the ectoparasitic mites are considered as major threats to honey bee colonies and health. However, their general biology remains poorly understood. Results We sequenced the genome and transcriptomes of Tropilaelaps mercedesae, the prevalent ectoparasitic mite infesting honey bees in Asia. The de novo assembled genome sequence (353 Mb) represents 53% of the estimated genome size because of the compression of repetitive sequences; nevertheless, we predicted 15,190 protein-coding genes which were well supported by the mite transcriptomes and proteomic dataes. Although amino acid substitutions have been accelerated within the conserved core genes in of two mites, T. mercedesae and Metaseiulus occidentalis, T. mercedesae has undergone the least gene family expansion and contraction between the seven arthropods we tested. The number of sensory system genes has been dramatically reduced; meanwhile, T. mercedesae may have evolved a specialized cuticle and water homeostasis mechanisms, as well as epigenetic control of gene expression for ploidy compensation between males and females., and water homeostasis. T. mercedesae contains all gene sets required to detoxify xenobiotics, enabling it to be miticide resistant. T. mercedesae is closely associated with a symbiotic bacteriuma (Rickettsiella grylli-like) and DWVdeformed wing virus (DWV), the most prevalent honey bee virus. The presence of DWV in both adult male and female mites was also confirmed by the proteomic analysis. Conclusions T. mercedesae has a very specialized life history and habitat as the ectoparasitic mite strictly dependsing on the honey bee inside the a stable colony. Thus, comparison of the genome and transcriptome sequences with those of a tick and free-living mites and tick has revealed the specific features of the genome shaped by interaction with the honey bee and colony environment. T. mercedesae, as well as Varroa destructor, genome and transcriptome sequences not only provide insights into the mite biology, but may also help to develop measures to control the most serious pests of the honey bee.
Project description:Schizosaccharomyces pombe is a model unicellular eukaryote with ties to the basic research, oenology and industrial biotechnology sectors. While most investigations into S. pombe cell biology utilize Leupold’s 972h- laboratory strain background, recent studies have described a wealth of genetic and phenotypic diversity within wild populations of S. pombe including stress resistance phenotypes which may be of interest to industry. Here we describe the genomic and transcriptomic characterization of Wilmar-P, an S. pombe isolate used for bioethanol production from sugarcane molasses at industrial scale. Novel sequences present in Wilmar-P but not in the laboratory S. pombe genome included multiple coding sequences with near-perfect nucleotide identity to Schizosaccharomyces octosporus sequences. Wilmar-P also contained a ~100kb duplication in the right arm of chromosome III, a region harboring ght5+, the predominant hexose transporter encoding gene. Transcriptomic analysis of Wilmar-P grown in molasses revealed strong downregulation of core environmental stress response genes and upregulation of hexose transporters and drug efflux pumps compared to laboratory S. pombe. Finally, examination of the regulatory network of Scr1, which is involved in the regulation of several genes differentially expressed on molasses revealed expanded binding of this transcription factor in Wilmar-P compared to laboratory S. pombe in the molasses condition. Together our results point to both genomic plasticity and transcriptomic adaptation as mechanisms driving phenotypic adaptation of Wilmar-P to the molasses environment and therefore adds to our understanding of genetic diversity within industrial fission yeast strains and the capacity of this strain for commercial scale bioethanol production.
Project description:Purpose : The goal of this study was to use RNA Seq to explore the correlation of gene expression of a collection of clinical P. aeruginosa isolates to various phenotypes, such as colony morphology, c-di-GMP production and biofilm formation Methods : mRNA profiles were generated for Pseudomonas aerugionsa clinical samples by deep sequencing.Ribosomal RNA was removed by using the RiboZero Bacteria kit (Illumina). cDNA libraries were synthesized using the SMARTScribe Reverse Transcriptase (Takara) followed by a PCR enrichment using the AccuPrime HiFi Taq polymerase (Invitrogen). Enzymatic reactions were carried out in the presence of SUPERase·In™ RNase Inhibitor (Invitrogen); RNACleanXP beads (Agencourt) were used for all RNA purification steps. Quality checks were performed before, during and after cDNA library preparation with the RNA Nano Kit and an Agilent Bioanalyzer 2100 (Agilent Technologies). Libraries were sequenced on an Illumina NovaSeq 6000 (paired-end mode; 2 x 50 bp) and mRNA reads were trimmed using the tool ‘cutadapt’ (version 3.5) with customized settings and mapped to the NC_008463.1 (PA14) reference genome from NCBI using ‘bowtie2’ (version 2.3.5.1) with the settings “--very-sensitive-local; --no-mixed; --fr; --no-unal”.